The ClpXP protease is dispensable for degradation of unfolded proteins in Staphylococcus aureus

Steen G. Stahlhut, Abdulelah A. Alqarzaee, Camilla Jensen, Niclas S. Fisker, Ana R. Pereira, Mariana G. Pinho, Vinai Chittezham Thomas, Dorte Frees

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

In living cells intracellular proteolysis is crucial for protein homeostasis, and ClpP proteases are conserved between eubacteria and the organelles of eukaryotic cells. In Staphylococcus aureus, ClpP associates to the substrate specificity factors, ClpX and ClpC forming two ClpP proteases, ClpXP and ClpCP. To address how individual ClpP proteases impact cell physiology, we constructed a S. aureus mutant expressing ClpX with an I265E substitution in the ClpP recognition tripeptide of ClpX. This mutant cannot degrade established ClpXP substrates confirming that the introduced amino acid substitution abolishes ClpXP activity. Phenotypic characterization of this mutant showed that ClpXP activity controls cell size and is required for growth at low temperature. Cells expressing the ClpXI265E variant, in contrast to cells lacking ClpP, are not sensitive to heat-stress and do not accumulate protein aggregates showing that ClpXP is dispensable for degradation of unfolded proteins in S. aureus. Consistent with this finding, transcriptomic profiling revealed strong induction of genes responding to protein folding stress in cells devoid of ClpP, but not in cells lacking only ClpXP. In the latter cells, highly upregulated loci include the urease operon, the pyrimidine biosynthesis operon, the betA-betB operon, and the pathogenicity island, SaPI5, while virulence genes were dramatically down-regulated.

Original languageEnglish (US)
Article number11739
JournalScientific reports
Volume7
Issue number1
DOIs
StatePublished - Dec 1 2017

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Protein Unfolding
Proteolysis
Staphylococcus aureus
Peptide Hydrolases
Operon
Genomic Islands
Cell Physiological Phenomena
Urease
Protein S
Protein Folding
Eukaryotic Cells
Amino Acid Substitution
Substrate Specificity
Cell Size
Organelles
Genes
Virulence
Homeostasis
Hot Temperature
Bacteria

ASJC Scopus subject areas

  • General

Cite this

Stahlhut, S. G., Alqarzaee, A. A., Jensen, C., Fisker, N. S., Pereira, A. R., Pinho, M. G., ... Frees, D. (2017). The ClpXP protease is dispensable for degradation of unfolded proteins in Staphylococcus aureus. Scientific reports, 7(1), [11739]. https://doi.org/10.1038/s41598-017-12122-y

The ClpXP protease is dispensable for degradation of unfolded proteins in Staphylococcus aureus. / Stahlhut, Steen G.; Alqarzaee, Abdulelah A.; Jensen, Camilla; Fisker, Niclas S.; Pereira, Ana R.; Pinho, Mariana G.; Thomas, Vinai Chittezham; Frees, Dorte.

In: Scientific reports, Vol. 7, No. 1, 11739, 01.12.2017.

Research output: Contribution to journalArticle

Stahlhut, SG, Alqarzaee, AA, Jensen, C, Fisker, NS, Pereira, AR, Pinho, MG, Thomas, VC & Frees, D 2017, 'The ClpXP protease is dispensable for degradation of unfolded proteins in Staphylococcus aureus', Scientific reports, vol. 7, no. 1, 11739. https://doi.org/10.1038/s41598-017-12122-y
Stahlhut SG, Alqarzaee AA, Jensen C, Fisker NS, Pereira AR, Pinho MG et al. The ClpXP protease is dispensable for degradation of unfolded proteins in Staphylococcus aureus. Scientific reports. 2017 Dec 1;7(1). 11739. https://doi.org/10.1038/s41598-017-12122-y
Stahlhut, Steen G. ; Alqarzaee, Abdulelah A. ; Jensen, Camilla ; Fisker, Niclas S. ; Pereira, Ana R. ; Pinho, Mariana G. ; Thomas, Vinai Chittezham ; Frees, Dorte. / The ClpXP protease is dispensable for degradation of unfolded proteins in Staphylococcus aureus. In: Scientific reports. 2017 ; Vol. 7, No. 1.
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